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1.
Clin Ter ; 173(2): 149-154, 2022 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-35385038

RESUMO

Objectives: Proximal femur fractures (PFF) usually occur in ge-riatric patients and lead to high mortality and disability. This study aims to analyze the mortality rate at 30 days in over-65 -years hospitalized patients with proximal femur fractures, comparing the data collected from three public hospitals in Lazio Region, focusing on early surgery outcomes. Materials and Methods: They were gathered data from the Regional Program for the Evaluation of the Outcomes of Health Interventions (P.Re.Val.E.) dated 2019, managed by the Department of Epidemiology of the Regional Health Service of Lazio (DEP), concerning femur fractures' treatment and outcome in elderly patients. Three hospitals were selected (called A, B, or C), belonging to a specific Local He-alth Authority in Rome, and for each of them were identified some indicators (total number of hospitalizations, origin of the admissions, destination of patients, surgery within 48 hours or two days, 30-day mortality, number of hospitalizations). Results: In Hospital A, the mortality rate at 30 days (305 registe-red patients) was 2.6%, the lowest among the hospitals considered. In Hospital B, the total mortality at 30 days (254 registered patients) was 7.48%, above the regional average of 5.95%. In Hospital C, the mortality rate at 30 days (71 registered patients) was 4.23%, below the regional average. Conclusions: P.Re.Val.E. represents a fundamental tool to evaluate the work of the Local Health Authorities and the structures that are part of it. In particular, about proximal femoral fractures in over-65-years patients, the results of surgical treatment in 48 hours and the morta-lity rate at 30 days represent a reproducible index of quality of the healthcare system. However, other variables need to be considered in future studies, taking into account different features of various hospitals, even if they belong to the same Local Health Authority. v.


Assuntos
Fraturas do Fêmur , Fraturas do Quadril , Idoso , Atenção à Saúde , Fraturas do Fêmur/cirurgia , Fêmur , Fraturas do Quadril/cirurgia , Hospitalização , Hospitais , Humanos , Estudos Retrospectivos
2.
Clin Ter ; 173(1): 1-5, 2022 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-35147638

RESUMO

ABSTRACT: Post-traumatic gas gangrene is a rare but potentially life-threa-tening condition due to soft tissues infection by Clostridium species. These anaerobic microaerophile bacteria are highly resistant to external insults related to their ability to produce spores, which can survive on any surface for long periods. Under certain conditions suitable for proliferation (such as in ischemic tissues), bacteria produce many to-xins. In particular, Clostridium perfringens type A represents the most frequent cause of traumatic gas gangrene nowadays. It produces toxins responsible for thrombotic and necrotic phenomena in soft tissues and rapid disease diffusion to muscles. Clinical manifestations usually start as local edema and emphysema but rapidly evolve into a septic state. Prognosis is poor in 20-30% of cases, and death occurs due to multiorgan failure. Because of its rapid evolution, clinical diagnosis is not always obtained, thus determining the need for post-mortem investigation. This case report presents a rare case of fulminant gas gangrene due to Clostridium Perfrigens infection developed after trau-matic injury. Despite the prompt antibiotic administration and surgical intervention on the site of trauma, gas gangrene rapidly evolved into septic shock, leading to the patient's death. Post-mortem investigations were conducted and confirmed multiorgan failure as the cause of death. Cultural analysis was also performed but showed no bacterial growth. Negativity on culture tests should be related to antibiotic administration before blood sampling and bacterial characteristics. In such cases, the correct identification of the cause of death was only possible following a careful and detailed forensic methodological approach.


Assuntos
Infecções por Clostridium , Gangrena Gasosa , Choque Séptico , Infecções por Clostridium/complicações , Infecções por Clostridium/diagnóstico , Clostridium perfringens , Gangrena Gasosa/diagnóstico , Gangrena Gasosa/etiologia , Humanos , Necrose
3.
Clin Ter ; 172(4): 369-371, 2021 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-34247221

RESUMO

ABSTRACT: Anaphylaxis is defined as a rapid systemic reaction that develops in individuals previously exposed to specific allergens. The new exposure causes systemic cellular degranulation, which in turn leads to cardiovascular and respiratory changes that are fatal if not treated immediately. One of the main problems of this scenario in the forensic field is the determination of a correct post-mortem diagnosis. Traditional methods, such as histopathological examination of the respiratory tract and the use of specific antibodies used in immunohistochemistry, are sensitive but not always specific and therefore do not guarantee a high degree of probability in the diagnosis of anaphylaxis. For this reason, a new and promising research frontier in this field of forensic pathology could be represented by the application of miRNAs as biomarkers, as has been done in other areas of medicine.


Assuntos
Anafilaxia/diagnóstico , Anafilaxia/mortalidade , Anafilaxia/fisiopatologia , Autopsia/métodos , Causas de Morte , Patologia Legal/métodos , Biomarcadores/análise , Humanos , Triptases/análise
4.
Eat Weight Disord ; 24(1): 3-11, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28290118

RESUMO

PURPOSE: The differential prevalence of eating disorders in males and females can be explained by the impact of gender-role orientations. Inside the Italian socio-cultural context, gender socialization can be influenced by stereotypical gender beliefs, and this may contribute to the psychological distress of individuals who identify with discrepant gender roles from their biological sex. Our study explored, within the Italian context, the potential moderating effect of masculinity and femininity on the relationships between gender and attitudes about body and eating. METHODS: Nine hundred and twenty Italian male and female adolescents (M = 427, F = 493; age 14-21 years) completed the Eating Disorder Inventory-2 (EDI-2) and the Bem Sex-Role Inventory (BSRI). RESULTS: A moderating effect of gender role on the relationship between gender and bulimia, and drive of thinness emerged. Girls with higher levels of masculinity scored higher on bulimia than did their counterparts with lower levels, and boys with higher levels of femininity scored higher on bulimia and on drive for thinness than did their counterparts with lower levels. Data did not reveal a moderating effect of gender role on the relationship between gender and body satisfaction. CONCLUSIONS: Our data suggest that adolescents who endorsed a gender role that is socially considered discrepant from their biological sex (girls with higher levels of masculinity and boys with higher levels of femininity) are more likely to show higher level of bulimia and drive of thinness. This suggests the need for prevention and treatment programmes for eating disorders that take into account individuals' gender-role orientation and the influence that culturally dominant gender beliefs can exert on it.


Assuntos
Atitude , Imagem Corporal/psicologia , Cultura , Ingestão de Alimentos/psicologia , Comportamento Alimentar/psicologia , Identidade de Gênero , Adolescente , Bulimia/psicologia , Feminino , Humanos , Itália , Masculino , Motivação , Satisfação Pessoal , Fatores Sexuais , Magreza/psicologia , Adulto Jovem
6.
Bull Environ Contam Toxicol ; 100(2): 240-244, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29181607

RESUMO

Cryptosporidium spp. has been recognized as an important pathogen. As bivalve mollusks are noted as potential sources of several pathogens due to their consumption as foodstuffs, the aim of this study was to investigate the occurrence of Cryptosporidium spp. oocysts in Perna perna mussels and in seawater samples from a mussel farm in Southeastern Brazil, where mussels are grown directly in the sea, attached to ropes. Oocysts were observed by microscopy and confirmed by an enzyme-linked immunosorbent assay. Oocysts were present in mussel gills and GI tracts, as well as in the seawater. Of the 100 females, 10% and 11% showed contaminated GI tracts and gills, respectively, while this rate was lower in males, at 5% and 8.9%. Oocysts were present in higher amounts in the GI tract compared to gills and water. Contamination of the study area is apparent, leading to public health risks. More in-depth studies are needed, including molecular investigations, to identify Cryptosporidium species in mussels, as well as the implementation of monitoring actions in animals destined for human consumption.


Assuntos
Cryptosporidium/fisiologia , Exposição Dietética/estatística & dados numéricos , Monitoramento Ambiental , Contaminação de Alimentos/estatística & dados numéricos , Perna (Organismo)/patogenicidade , Frutos do Mar/estatística & dados numéricos , Animais , Brasil , Brânquias , Humanos , Oocistos , Alimentos Marinhos , Água do Mar
7.
Eur Psychiatry ; 40: 65-75, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27992836

RESUMO

BACKGROUND: Subjects at ultra high-risk (UHR) for psychosis have an enhanced vulnerability to develop the disorder but the risk factors accounting for this accrued risk are undetermined. METHOD: Systematic review of associations between genetic or environmental risk factors for psychosis that are widely established in the literature and UHR state, based on comparisons to controls. RESULTS: Forty-four studies encompassing 170 independent datasets and 54 risk factors were included. There were no studies on association between genetic or epigenetic risk factors and the UHR state that met the inclusion criteria. UHR subjects were more likely to show obstetric complications, tobacco use, physical inactivity, childhood trauma/emotional abuse/physical neglect, high perceived stress, childhood and adolescent low functioning, affective comorbidities, male gender, single status, unemployment and low educational level as compared to controls. CONCLUSIONS: The increased vulnerability of UHR subjects can be related to environmental risk factors like childhood trauma, adverse life events and affective dysfunction. The role of genetic and epigenetic risk factors awaits clarification.


Assuntos
Exposição Ambiental/estatística & dados numéricos , Transtornos Psicóticos/epidemiologia , Transtornos Psicóticos/psicologia , Meio Social , Adolescente , Meio Ambiente , Feminino , Humanos , Masculino , Escalas de Graduação Psiquiátrica , Fatores de Risco , Ajustamento Social
8.
J Mater Sci Mater Med ; 25(9): 2205-14, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24997984

RESUMO

The infections give rise to a range of clinical problems and prolong hospitalization with increased healthcare costs. Moreover, persistent infections exasperate the problem of antibiotic resistance. The aim of this study was the development of effective and low-cost antibacterial silver coatings on surgical sutures by adopting an innovative photochemical deposition process to prevent early contamination of surgical wounds. The silver deposition technology adopted in this work is an innovative process based on the in situ photoreduction of a silver solution. The samples were dipped in the silver solution and then exposed to UV radiation in order to induce the synthesis of silver clusters on the surface of the suture. The homogeneous distribution of silver particles on the surface and on the cross-section of the treated sutures was demonstrated. All the antibacterial studies clearly demonstrated that the use of novel silver treated sutures could represent clinical advantages in terms of the prevention of surgical infections against bacterial colonization. The silver coating deposited on the sutures demonstrated no cytotoxic effect on a selected cell population. The results obtained suggested that the antibacterial silver-coated sutures developed in this work could represent an interesting alternative to conventional sutures, with evident advantages in terms of prevention of the surgical infections and on the health costs. In addiction, very low concentrations of silver significantly inhibited the microbial load, without affecting the cell viability.


Assuntos
Materiais Revestidos Biocompatíveis , Nanopartículas Metálicas , Seda , Prata/química , Infecção da Ferida Cirúrgica/prevenção & controle , Suturas/efeitos adversos , Células 3T3 , Animais , Antibacterianos/administração & dosagem , Antibacterianos/farmacologia , Contagem de Colônia Microbiana , Escherichia coli/efeitos dos fármacos , Escherichia coli/isolamento & purificação , Camundongos , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/isolamento & purificação , Resistência à Tração
9.
Minerva Chir ; 67(5): 429-32, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23232481

RESUMO

AIM: In the evaluation of nodular thyroid lesions, cytology is the main diagnostic instrument, associated with ultrasound examination. METHODS: We prospectively categorized into a dedicated database, 612 patients submitted to thyroidectomy from January 2009 to December 2011. We selected two groups of patients: the cases with "follicular lesions" and the cases classified as "THYR 3" using Bethesda Classification. RESULTS: Of 612 patients submitted to TT, in 68 cases, 8 males (M) and 60 females (F), we recorded a preoperative cytological diagnosis compatible with THYR 3 class. In 56 cases, 82.4% of the patients (5 M, 51 F), we performed a thyroidectomy with Conventional technique (CT) and in 12 cases, 12.6% of the patients (3M, 9F) with video-assisted technique (MIVAT). In 39 cases (57.1%) of the THYR 3 patients treated, a benign disease was found while in the other 29 cases (42.9%) a differentiated thyroid cancer (DTC) were found. In patients with nodules classified as "follicular proliferation", treated in the same period and by the same surgical team, we recorded 97 cases (79 F, 18 M). The patients were treated with TT in 72 cases, with MIVAT in 22 cases; in 3 cases we performed a HT with traditional technique. Adenomatous hyperplasia was found in 51; DTC was found in 21 cases and adenoma in 25 cases. The analysis of the incidence of benign versus malignant disease between the 2 groups showed a P<0.0001. CONCLUSION: In our experience, we recommend surgery, TT or HT, for THYR 3 nodules analyzed for the first time by FNAB with sonographic pattern suspicious for malignancy.


Assuntos
Doenças da Glândula Tireoide/patologia , Doenças da Glândula Tireoide/cirurgia , Neoplasias da Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/cirurgia , Tireoidectomia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Cuidados Pré-Operatórios , Estudos Prospectivos , Doenças da Glândula Tireoide/classificação , Neoplasias da Glândula Tireoide/classificação
10.
Braz. j. morphol. sci ; 28(1): 4-10, Jan.-Mar. 2011. ilus
Artigo em Inglês | LILACS | ID: lil-644124

RESUMO

Embryos of Caiman yacare were collected and subjected to the bone clearing and staining protocol in orderto analyze the ontogenetic patterns of ossification of the pectoral girdle and forelimb skeleton. The osseousstructure of the girdle and forelimbs of C. yacare begins to ossify starting at 30 days of incubation, withthe presence of dye retention in the scapula, coracoids, humerus, radius and ulna bones. During embryonicdevelopment, the autopodio of C. yacare has four bones in the carpus, the radial, ulnar, pisiform and carpaldistal 4+5 bone. Their ossification begins at 39 days of incubation with the radial, followed by the ulnar, and at54 days, the pisiform and the distal carpal 4 + 5. Each mesopodio has 5 metacarpi and are present 15 phalanges,two in digits I and V, three in digits II and IV, and four in digit III (phalangeal formula 2:3:4:3:2). Ossificationof the metacarpi starts at 27 days of incubation, following the sequence MCII=MCIII=MCIV>MCI>MCV.The first phalanges begin the process of ossification on day 36, continuing up to the last day of incubation.The sequence of ossification of the proximal phalanges is PPI=PPII=PPIII>PPIV=PPV, that of the medialphalanges is MPII>MPpIII>MPdIII>MPIV, and that of the distal phalanges is DPI>DPII>DPIII>DPV>DPIV.The ontogenetic pattern of the bones of the forepaw of C. yacare generally differs from that of other reptiles,although there are some similarities.


Assuntos
Animais , Membro Anterior/anatomia & histologia , Membro Anterior/embriologia , Osteogênese/fisiologia , Osteogênese/genética , Jacarés e Crocodilos/fisiologia , Jacarés e Crocodilos/metabolismo , Répteis
11.
Vaccine ; 29(9): 1801-11, 2011 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-21215342

RESUMO

The Plasmodium vivax Merozoite Surface Protein-3α (PvMSP-3α) is considered as a potential vaccine candidate. However, the detailed investigations of the type of immune responses induced in naturally exposed populations are necessary. Therefore, we aim to characterize the naturally induced antibody to PvMSP-3α in 282 individuals with different levels of exposure to malaria infections residents in Brazilian Amazon. PvMSP3 specific antibodies (IgA, IgG and IgG subclass) to five recombinant proteins and the epitope mapping by Spot-synthesis technique to full-protein sequence of amino acids (15aa sequence with overlapping sequence of 9aa) were performed. Our results indicates that PvMSP3 is highly immunogenic in naturally exposed populations, where 78% of studied individuals present IgG immune response against the full-length recombinant protein (PVMSP3-FL) and IgG subclass profile was similar to all five recombinant proteins studied with a high predominance of IgG1 and IgG3. We also observe that IgG and subclass levels against PvMSP3 are associated with malaria exposure. The PvMSP3 epitope mapping by Spot-synthesis shows a natural recognition of at least 15 antigenic determinants, located mainly in the two blocks of repeats, confirming the high immunogenicity of this region. In conclusion, PvMSP-3α is immunogenic in naturally exposed individuals to malaria infections and that antibodies to PvMSP3 are induced to several B cell epitopes. The presence of PvMSP3 cytophilic antibodies (IgG1 and IgG3), suggests that this mechanism could also occur in P. vivax.


Assuntos
Anticorpos Antiprotozoários/química , Antígenos de Protozoários/imunologia , Mapeamento de Epitopos/métodos , Epitopos de Linfócito B/imunologia , Malária Vivax/imunologia , Plasmodium vivax/imunologia , Proteínas de Protozoários/imunologia , Adulto , Sequência de Aminoácidos , Anticorpos Antiprotozoários/genética , Antígenos de Protozoários/genética , Brasil/epidemiologia , Estudos de Coortes , Estudos Transversais , Epitopos de Linfócito B/genética , Feminino , Humanos , Malária Vivax/epidemiologia , Malária Vivax/genética , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Plasmodium vivax/genética , Proteínas de Protozoários/genética , Adulto Jovem
12.
Vaccine ; 28(18): 3185-91, 2010 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-20189487

RESUMO

Plasmodium vivax merozoite surface protein (PvMSP9) stimulates both cellular and humoral immune responses in individuals who are naturally infected by this parasite species. To identify immunodominant human T-cell epitopes in PvMSP9, we used the MHC class II binding peptide prediction algorithm ProPred. Eleven synthetic peptides representing predicted putative promiscuous T-cell epitopes were tested in IFN-gamma and IL-4 ELISPOT assays using peripheral blood mononuclear cells (PBMC) derived from 142 individuals from Rondonia State, Brazil who had been naturally exposed to P. vivax infections. To determine whether the predicted epitopes are preferentially recognized in the context of multiple alleles, MHC Class II typing of the cohort was also performed. Five synthetic peptides elicited robust cellular responses, and the overall frequencies of IFN-gamma and IL-4 responders to at least one of the promiscuous peptides were 62% and 46%, respectively. The frequencies of IFN-gamma and IL-4 responders to each peptide were not associated with a particular HLA-DRB1 allelic group since most of the peptides induced a response in individuals of 12 out of 13 studied allelic groups. The prediction of promiscuous epitopes using ProPred led to the identification of immunodominant epitopes recognized by PBMC from a significant proportion of a genetically heterogeneous population exposed to malaria infections. The combination of several such T-cell epitopes in a vaccine construct may increase the frequency of responders and the overall efficacy of subunit vaccines in genetically distinct populations.


Assuntos
Epitopos de Linfócito T/imunologia , Interferon gama/metabolismo , Interleucina-4/metabolismo , Leucócitos Mononucleares/imunologia , Malária Vivax/imunologia , Proteínas de Membrana/imunologia , Plasmodium vivax/imunologia , Proteínas de Protozoários/imunologia , Adulto , Alelos , Animais , Brasil , Mapeamento de Epitopos , Antígenos HLA-DR/genética , Cadeias HLA-DRB1 , Humanos , Adulto Jovem
13.
J Endocrinol Invest ; 32(7): 601-5, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19498323

RESUMO

AIMS: The aim of this study was to establish whether short-term GH treatment causes obstructive apnea in patients with Prader-Willi syndrome and normal upper airway patency. SUBJECTS AND METHODS: We performed an observational longitudinal 6-week GH treatment study. Thirty-four non-severely obese Prader-Willi syndrome patients (20 boys, age range 0.94-11.8 yr, median 2.24 yr) entered an observational longitudinal 6-week study. Sixteen boys received recombinant human GH (rhGH) treatment; the remaining 18 represented the control group and received no treatment. Polysomnography monitoring and othorhinolaringoiatric video endoscopy were performed one night before and after 6 weeks of rhGH treatment (0.03 mg/kg body weight/day). All patients underwent auxologic assessment, fasting blood glucose, insulin and IGF-I evaluation. The main polysomnographic parameter considered was total apnea hypopnea index, consisting of two components: central apnea hypopnea index and obstructive apnea hypopnea index. All patients were free of severe or moderate upper airway obstruction when rhGH treatment began. RESULTS: After 6 weeks of rhGH therapy, obstructive apnea hypopnea index increased in 8/16 (50%), decreased in 5/16 (31%), and did not change in 3/16 (19%) patients. The changes were not statistically significant. The rhGH-treated group did not differ from the control group for the apnea hypopnea index both before and after 6 weeks of treatment. Adenoids and tonsils showed a slight increase in 1 and 2 patients on rhGH treatment, respectively, and did not change in the untreated patients. CONCLUSIONS: Our data show that short-term rhGH treatment does not cause restrictions of the upper airways in patients with Prader-Willi syndrome and normal upper airway patency.


Assuntos
Hormônio do Crescimento Humano/uso terapêutico , Obesidade/complicações , Síndrome de Prader-Willi , Proteínas Recombinantes/uso terapêutico , Apneia Obstrutiva do Sono , Traqueia/efeitos dos fármacos , Antropometria , Glicemia/metabolismo , Composição Corporal , Índice de Massa Corporal , Criança , Pré-Escolar , Humanos , Lactente , Insulina/sangue , Resistência à Insulina/fisiologia , Masculino , Polissonografia , Síndrome de Prader-Willi/tratamento farmacológico , Síndrome de Prader-Willi/fisiopatologia , Apneia Obstrutiva do Sono/etiologia , Apneia Obstrutiva do Sono/fisiopatologia , Apneia Obstrutiva do Sono/terapia , Traqueia/patologia
14.
Toxicon ; 52(8): 944-53, 2008 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-18948130

RESUMO

BJ-32 (also known as BjcuL) is a C-type lectin from the venom of Bothrops jararacussu with specificity for beta-galactosides and a remarkable ability to agglutinate several species of trypanosomatids. Our objective was to study the oligomerization state of native BJ-32 by using different biophysical and computational methods. Small-angle X-ray light scattering (SAXS) experiments disclosed a compact, globular protein with a radius of gyration of 36.72+/-0.04A and molecular weight calculated as 147.5+/-2.0kDa. From analytical ultracentrifugation analysis, it was determined that the BJ-32 sedimentation profile fits nicely to a decamer model. The analysis of the intrinsic emitted fluorescence spectra for BJ-32 solutions indicated that association of subunits in the decamer is accompanied by changes in the environment of Tryptophan residues. Both ab initio and comparative models of BJ-32 supported the resemblance of the decamer in the crystallographic structure from a close homologue, the rattlesnake venom lectin (RSL) from Crotalus atrox.


Assuntos
Bothrops , Venenos de Crotalídeos/química , Lectinas Tipo C/química , Sequência de Aminoácidos , Animais , Dicroísmo Circular , Simulação por Computador , Venenos de Crotalídeos/análise , Venenos de Crotalídeos/isolamento & purificação , Lectinas Tipo C/análise , Lectinas Tipo C/isolamento & purificação , Modelos Moleculares , Dados de Sequência Molecular , Estrutura Quaternária de Proteína , Espalhamento a Baixo Ângulo , Alinhamento de Sequência , Espectrometria de Massas por Ionização por Electrospray , Ultracentrifugação
15.
Vaccine ; 26(51): 6645-54, 2008 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-18832003

RESUMO

Antibody and T-cell reactivities to Plasmodium vivax merozoite surface protein 9 (PvMSP9) were evaluated in a cross-sectional study of individuals naturally exposed to malaria infections living in Ribeirinha, a native riverine community and in Colina, a transmigrant community, Rondonia, Brazil. The antibody responses to PvMSP9-RIRIIand PvMSP9-Nt domains in Ribeirinha were higher compared with Colina and correlated with age and time of malaria exposure. IgG2 was most prevalent for PvMSP9-RII in both communities, and IgG1 was the predominant isotype for PvMSP9-Nt and PvMSP9-RIRII in Ribeirinha. IFN-gamma and IL-4 predominated in Ribeirinha, while IFN-gamma predominated in Colina. Variation in exposure to P. vivax likely accounts for the differences observed in cytokine and antibody levels between the two populations studied.


Assuntos
Anticorpos Antiprotozoários/imunologia , Malária Vivax/imunologia , Proteínas de Membrana/imunologia , Plasmodium vivax/imunologia , Proteínas de Protozoários/imunologia , Adulto , Animais , Anticorpos Antiprotozoários/sangue , Formação de Anticorpos/imunologia , Brasil/epidemiologia , Estudos de Coortes , Estudos Transversais , Epitopos de Linfócito T/imunologia , Feminino , Humanos , Imunidade Ativa , Imunidade Celular , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Interferon gama/sangue , Interferon gama/imunologia , Interleucina-4/sangue , Interleucina-4/imunologia , Malária Vivax/epidemiologia , Masculino , Pessoa de Meia-Idade , Prevalência , Proteínas Recombinantes/imunologia , Adulto Jovem
16.
Parasitol Res ; 101(6): 1627-35, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17726617

RESUMO

To investigate the importance of serine proteases in Leishmania amazonensis promastigotes, we analyzed the effects of classical serine protease inhibitors and a Kunitz-type inhibitor, obtained from sea anemone Stichodactyla helianthus (ShPI-I), on the viability and morphology of parasites in culture. Classical inhibitors were selected on the basis of their ability to inhibit L. amazonensis serine proteases, previously described. The N-tosyl-L: -phenylalanine chloromethyl ketone (TPCK) and benzamidine (Bza) inhibitors, which are potential Leishmania proteases inhibitors, in all experimental conditions reduced the parasite viability, with regard to time dependence. On the other hand, N-tosyl-lysine chloromethyl ketone (TLCK) did not significantly affect the parasite viability, as it was poor Leishmania enzymes inhibitor. Ultrastructural analysis demonstrated that both Bza and TPCK induced changes in the flagellar pocket region with membrane alteration, including bleb formation. However, TPCK effects were more pronounced than those of Bza in Leishmania flagellar pocket in plasma membrane, and intracellular vesicular bodies was visualized. ShPI-I proved to be a powerful inhibitor of L. amazonensis serine proteases and the parasite viability. The ultrastructural alterations caused by ShPI-I were more dramatic than those induced by the classical inhibitors. Vesiculation of the flagellar pocket membrane, the appearance of a cytoplasmic vesicle that resembles an autophagic vacuole, and alterations of promastigotes shape resulted.


Assuntos
Leishmania mexicana/efeitos dos fármacos , Leishmania mexicana/crescimento & desenvolvimento , Serina Endopeptidases/metabolismo , Inibidores de Serina Proteinase/farmacologia , Animais , Colorimetria , Leishmania mexicana/enzimologia , Leishmania mexicana/ultraestrutura , Microscopia Eletrônica , Testes de Sensibilidade Parasitária , Sais de Tetrazólio , Tiazóis
17.
Artigo em Inglês | MEDLINE | ID: mdl-16750943

RESUMO

Affinity chromatography based on the commercial resin Sepharose CL-6B was used to isolate new C1-beta-type lectins from crude preparations of snake venoms (Bothrops jararaca, Bothrops jararacussu, Bothrops newiedi, Bothrops moojeni, Lachesis muta rhombeata). Most of the C-type lectins could be eluted with almost 100% recovery using the competitor isopropyl-beta-D-thiogalactoside (IPTG) or through Ca2+ sequestration with EDTA. The lectin yield varied considerably among the different snake species, but B. newiedi venom was a particularly rich source of lectin, retaining 2.7 mg of lectin by milliliter of resin in saturating conditions. C1-alpha-lectins from Crotalus durisus terrificus venom, from the jack fruit (jacalin) and from bread fruit seeds extract (frutalin) had no affinity, either with or without Ca2+ added, for Sepharose CL-6B, showing that the resin is specific for C1-beta-type lectins. Sepharose CL-6B used as galactose-affinity chromatography provides a simple and fast method for isolating C-type beta-galactoside binding lectins from crude sample preparations.


Assuntos
Cromatografia de Afinidade/métodos , Galectinas/análise , Sequência de Aminoácidos , Animais , Bothrops/metabolismo , Venenos de Crotalídeos/análise , Venenos de Crotalídeos/química , Venenos de Crotalídeos/farmacologia , Galectinas/química , Galectinas/isolamento & purificação , Testes de Hemaglutinação , Humanos , Sefarose/análogos & derivados , Sefarose/química , Análise de Sequência de Proteína , Homologia de Sequência de Aminoácidos , Especificidade da Espécie , Venenos de Víboras/análise , Venenos de Víboras/química , Venenos de Víboras/farmacologia , Viperidae/metabolismo
18.
Biochimie ; 88(1): 59-68, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16085351

RESUMO

We show here that serum of piaussu, a Neotropical characin fish, has the highest butyrylcholinesterase activity so far described for humans and fish. To clarify whether this cholinesterase could protect piaussu against anticholinesterase pesticides by scavenging organophosphates, we purified it 1700-fold, with a yield of 80%. Augmenting concentrations (from 0.01 to 20 mM) of butyrylthiocholine activated it. The pure enzyme was highly inhibited by chlorpyriphos-oxon (ki=10,434x10(6) M-1 min-1) and by the specific butyrylcholinesterase inhibitor, isoOMPA (ki=45.7x10(6) M-1 min-1). Electrophoresis of total serum and 2-D electrophoresis of the purified cholinesterase showed that some enzyme molecules could circulate in piaussu serum as heterogeneously glycosylated dimers. The enzyme's N-terminal sequence was similar to sequences found for butyrylcholinesterase from sera of other vertebrates. Altogether, our data present a novel butyrylcholinesterase with the potential of protecting a fish from poisoning by organophosphates.


Assuntos
Butirilcolinesterase/sangue , Peixes/sangue , Sequência de Aminoácidos , Animais , Butirilcolinesterase/isolamento & purificação , Butirilcolinesterase/metabolismo , Butiriltiocolina/metabolismo , Clorpirifos/análogos & derivados , Clorpirifos/farmacologia , Inibidores da Colinesterase/farmacologia , Eletroforese em Gel Bidimensional , Eletroforese em Gel de Poliacrilamida , Humanos , Dados de Sequência Molecular , Intoxicação por Organofosfatos , Paraoxon/análogos & derivados , Paraoxon/farmacologia , Intoxicação/prevenção & controle , Alinhamento de Sequência , Tetraisopropilpirofosfamida/farmacologia
19.
Parasitology ; 131(Pt 1): 85-96, 2005 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16038400

RESUMO

A serine protease was purified 942-fold from culture supernatant of L. amazonensis promastigotes using (NH4)2SO4 precipitation followed by affinity chromatography on aprotinin-agarose and continuous elution electrophoresis by Prep Cell, yielding a total recovery of 61%. The molecular mass of the active enzyme estimated by SDS-PAGE under conditions of reduction was 56 kDa and 115 kDa under conditions of non-reduction, suggesting that the protease is a dimeric protein. Additionally, it was found to be a non-glycosylated enzyme, with a pI of 5.0. The optimal pH and temperature of the enzyme were 7.5 and 28 degrees C respectively, using alpha-N-rho-tosyl-L-arginine-methyl ester (L-TAME) as substrate. Assays of thermal stability indicated that 61% of the enzyme activity was preserved after 1 h of pre-treatment at 42 degrees C. Haemoglobin, bovine serum albumin (BSA), ovalbumin, fibrinogen, collagen, gelatin and peptide substrates containing arginine in an ester bond and amide substrates containing hydrophobic residues at the P1 site were hydrolysed by this extracellular protease. The insulin beta-chain was also hydrolysed by the enzyme and many peptidic bonds were susceptible to the protease action, and 4 of them (L11-V12, E3-A14, L15-Y16 and Y16-L17) were identified. Inhibition studies suggested that the enzyme belongs to the serine protease class inhibited by calcium and manganese and activated by zinc. These findings show that this enzyme of L. amazonensis is a novel serine protease, which differs from all known flagellate proteases characterized.


Assuntos
Leishmania/enzimologia , Serina Endopeptidases/metabolismo , Animais , Estabilidade Enzimática , Concentração de Íons de Hidrogênio , Inibidores de Proteases , Proteínas de Protozoários/metabolismo , Serina Endopeptidases/química , Especificidade por Substrato , Temperatura
20.
Artigo em Inglês | MEDLINE | ID: mdl-15994137

RESUMO

Snake venoms contain several trypsin-like enzymes with equivalent physicochemical characteristics and similar inhibition profiles. These are rather difficult to separate by classical purification procedures and therefore constitute a good model for affinity chromatography analysis. Some of these trypsin homologues present fibrinogenase activity, mimicking one or more features of the central mammalian coagulation enzyme, thrombin. It was previously demonstrated that a number of amidine derivatives are able to interact specifically with some of these serine proteases. To understand the enzyme-sorbent interactions we have investigated the ability of two commercially available benzamidine affinity matrices to purify thrombin-like serine proteases (TLSP) with similar biological properties from two snake venoms (Bothrops jararacussu and Lachesis muta rhombeata). Curiously, each sorbent retained a single but distinct TLSP from each venom with high yield. Molecular modeling analysis suggested that hydrophobic interactions within a specific region on the surface of these enzymes could be generated to explain this exquisite specificity. In addition, it was demonstrated that a specific tandem alignment of the two benzamidine sorbents enables the purification of three other enzymes from B. jararacussu venom.


Assuntos
Benzoilarginina Nitroanilida/química , Cromatografia de Afinidade/métodos , Venenos de Crotalídeos/enzimologia , Sefarose/análogos & derivados , Serina Endopeptidases/isolamento & purificação , Venenos de Víboras/enzimologia , Sequência de Aminoácidos , Animais , Bothrops , Interações Hidrofóbicas e Hidrofílicas , Modelos Moleculares , Sefarose/química , Serina Endopeptidases/metabolismo , Inibidores de Serina Proteinase/química , Viperidae
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